JP7768673B2 - Inspection device and inspection method - Google Patents

Description

The embodiments disclosed in this specification and drawings relate to an inspection device and an inspection method.

IVD (In Vitro Diagnostics) is a test that obtains indicators of a subject's condition from test samples such as blood, urine, or sputum. IVD detects target substances contained in the test sample, for example by detecting fluorescence. Specifically, a detection substance that specifically binds to a specific target substance is used in combination with a fluorescent dye, and based on the detection results of the fluorescence from the fluorescent dye, it can be determined whether the test sample contains a target substance that specifically binds to the detection substance.

Technology is known for simultaneously detecting multiple types of target substances and shortening testing time. For example, by assigning a fluorescent wavelength band to each target substance and detecting the fluorescence in each wavelength band, it is possible to simultaneously detect multiple types of target substances. However, depending on the number of target substances, the wavelength bands assigned to each target substance may become narrower, which may reduce the accuracy of detecting the fluorescence in each wavelength band.

JP 2010-261791 A

One of the problems that the embodiments disclosed in this specification and drawings attempt to solve is the accurate simultaneous detection of multiple types of target substances. However, the problems that the embodiments disclosed in this specification and drawings attempt to solve are not limited to the above problem. Problems corresponding to the effects of each configuration shown in the embodiments described below can also be positioned as other problems.

The testing device of this embodiment includes a detection unit and an identification unit. The detection unit detects, for each wavelength band, fluorescence emitted from a mixed sample prepared by mixing a sample with a reagent containing multiple combinations of fluorescent dyes and detection substances that specifically bind to specific target substances. The identification unit identifies the target substance contained in the sample based on the combination of multiple wavelength bands in which the fluorescence is detected.

FIG. 1 is a diagram showing an example of the configuration of an inspection apparatus according to the first embodiment. FIG. 2 is a flowchart showing an example of processing by the processing circuit of the inspection apparatus according to the first embodiment. FIG. 3 is a diagram showing an example of a reagent according to the first embodiment. FIG. 4 is a diagram showing an example of a magnetic particle according to the first embodiment. FIG. 5 is a diagram for explaining the fluorescence detection process for each wavelength band according to the first embodiment. FIG. 6 is a diagram showing an example of a fluorescence detection result according to the first embodiment. FIG. 7 is a diagram showing an example of a correspondence table between combinations of wavelength bands and standard substances according to the first embodiment. FIG. 8A is a diagram illustrating an example of a reaction vessel according to the second embodiment. FIG. 8B is a diagram showing an example of a reagent according to the second embodiment. FIG. 8C is a diagram showing an example of a reagent according to the second embodiment. FIG. 8D is a diagram showing an example of a reagent according to the second embodiment. FIG. 8E is a diagram showing an example of a fluorescence detection result according to the second embodiment. FIG. 9 is a diagram showing an example of a fluorescence detection result according to the third embodiment.

Embodiments of the inspection device and inspection method will be described in detail below with reference to the attached drawings.

(First embodiment)
1 is a diagram showing an example of the configuration of an inspection apparatus 1 according to the first embodiment. For example, the inspection apparatus 1 includes a console device 10, a dispensing control device 20, a detection device 30, a probe 40, and a reaction vessel 50. The console device 10 also includes a memory 11, an input interface 12, a display 13, and a processing circuit 14.

Memory 11 is realized, for example, by a semiconductor memory element such as RAM (Random Access Memory), flash memory, a hard disk, an optical disk, etc. For example, memory 11 stores programs that enable circuits included in inspection device 1 to realize their functions. Memory 11 also stores inspection results obtained by processing circuit 14, which will be described later. Memory 11 is an example of a storage unit.

The input interface 12 accepts various input operations from the user, converts the accepted input operations into electrical signals, and outputs them to the processing circuit 14. For example, the input interface 12 may be implemented by a mouse, keyboard, trackball, switch, button, joystick, touchpad that allows input operations by touching the operation surface, a touchscreen that integrates a display screen and touchpad, a non-contact input circuit using an optical sensor, a voice input circuit, etc. The input interface 12 may also be configured as a tablet terminal or the like that can wirelessly communicate with the console device 10. The input interface 12 may also be a circuit that accepts input operations from the user using motion capture. For example, the input interface 12 can process signals acquired via a tracker or images collected about the user to accept the user's body movements, line of sight, etc. as input operations. The input interface 12 is not limited to those equipped with physical operating components such as a mouse or keyboard. For example, an electrical signal processing circuit that receives electrical signals corresponding to input operations from an external input device provided separately from the console device 10 and outputs these electrical signals to the processing circuit 14 is also included as an example of the input interface 12.

The display 13 is a display device that displays various types of information. For example, the display 13 displays a GUI (Graphical User Interface) for receiving instructions from the user via the input interface 12. The display 13 also displays the test results from the processing circuitry 14, which will be described later. The display 13 may be implemented as a liquid crystal display, a CRT (Cathode Ray Tube) display, a touch panel, or the like.

The processing circuitry 14 controls the operation of the entire inspection device 1 by executing the control function 141, detection function 142, identification function 143, and output function 144. The control function 141 is an example of a control unit. The detection function 142 is an example of a detection unit. The identification function 143 is an example of an identification unit.

For example, the processing circuitry 14 controls the operation of the dispensing control device 20 and the detection device 30 (described below) by reading and executing a program corresponding to the control function 141 from the memory 11. For example, the control function 141 controls the operation of the dispensing control device 20 to mix the reagent and the test sample. The control function 141 also controls the operation of the detection device 30 to irradiate the mixed sample of the reagent and the test sample with light. The processing circuitry 14 also reads and executes a program corresponding to the detection function 142 from the memory 11 to detect fluorescence emitted from the mixed sample of the reagent and the test sample for each wavelength band. The processing circuitry 14 also reads and executes a program corresponding to the identification function 143 from the memory 11 to identify a target substance contained in the test sample based on a combination of multiple wavelength bands in which fluorescence was detected. The processing circuitry 14 also reads and executes a program corresponding to the output function 144 from the memory 11 to output the target substance identified by the identification function 143 as the test result. Details of each function of the processing circuitry 14 will be described later.

The dispensing control device 20 includes a drive mechanism 21 and a dispensing mechanism 22. The drive mechanism 21 drives the probe 40 and the reaction vessel 50. For example, the drive mechanism 21 adjusts the position of the probe 40 relative to the reagent vessel and the reaction vessel 50 so that the probe 40 can aspirate the reagent from the reagent vessel and inject the reagent into the reaction vessel 50. The dispensing mechanism 22 dispenses (aspirates and dispenses) the reagent from the probe 40 by, for example, using a syringe pump to aspirate water from the probe 40 and deliver water to the probe 40.

The detection device 30 includes a light source 31, a magnetic field generator 32, and a photodetector 33. The light source 31 irradiates the sample in the reaction vessel 50 with excitation light. For example, the light source 31 is configured by combining multiple light sources that emit light of different wavelengths as excitation light. As an example, the light source 31 is realized by a light-emitting diode (LED). The magnetic field generator 32 generates a magnetic field to be applied to the sample in the reaction vessel 50. The magnetic field generator 32 may be a temporary magnet such as an electromagnet, or a permanent magnet.

The photodetector 33 is a device that detects the fluorescence emitted from the sample in the reaction vessel 50 for each wavelength band. For example, the photodetector 33 can be realized by a detection device using a spectroscope and a photodetector, or a multi-spectral imaging (MSI) camera that captures two-dimensional images for each wavelength. The MSI camera divides the wavelength of light into multiple wavelength bands and acquires images for each wavelength band. Based on these images, it can be determined whether fluorescence in the wavelength band corresponding to each image has been detected.

The probe 40 is a device that injects reagents into the reaction vessel 50. For example, under the control of the dispensing control device 20, the probe 40 injects reagents containing multiple combinations of fluorescent dyes and detection substances that specifically bind to specific target substances into the reaction vessel 50, and mixes them with the test sample in the reaction vessel 50.

In FIG. 1, the testing device 1 is described as including a dispensing control device 20, a probe 40, and a reaction vessel 50, and as automatically injecting a sample into the reaction vessel 50, but this may be performed by the user. That is, the user may manually inject a reagent into the reaction vessel 50 by operating the probe 40 or reaction vessel 50. In this case, the testing device 1 may not include the dispensing control device 20, the probe 40, or the reaction vessel 50.

In the inspection device 1 shown in Figure 1, each processing function is stored in memory 11 in the form of a program that can be executed by a computer. The processing circuitry 14 is a processor that realizes the function corresponding to each program by reading and executing the program from memory 11. In other words, once a program has been read, the processing circuitry 14 has the function corresponding to the read program.

In FIG. 1, the control function 141, detection function 142, identification function 143, and output function 144 are described as being realized by a single processing circuit 14. However, the processing circuit 14 may also be configured by combining multiple independent processors, with each processor executing a program to realize the functions. Furthermore, each processing function possessed by the processing circuit 14 may be realized by being appropriately distributed or integrated into a single or multiple processing circuits. Furthermore, the processing circuit 14 may also realize its functions by using a processor in an external device connected via a network. For example, the processing circuit 14 realizes each function shown in FIG. 1 by reading and executing a program corresponding to each function from the memory 11, and by using a group of servers (cloud) connected to the inspection device 1 via a network as a computing resource.

The above describes an example configuration of the testing device 1 according to this embodiment. With this configuration, the processing circuitry 14 in the testing device 1 enables accurate simultaneous detection of multiple types of target substances.

The processing performed by the processing circuitry 14 will be described below with reference to the flowchart in Figure 2. Figure 2 is a flowchart showing an example of processing performed by the processing circuitry 14 of the inspection device 1 according to the first embodiment.

First, the control function 141 places the test sample P into the reaction vessel 50 (step S101). For example, the control function 141 causes the dispensing control device 20 to control the operation of the probe 40, thereby dispensing the test sample P from the probe 40 into the reaction vessel 50. Note that step S101 may also be performed manually by the user.

The test sample P is, for example, a biological sample such as blood, urine, or sputum. The testing device 1 can obtain an indicator of the subject's condition by testing whether the test sample P contains a specific target substance. The type of target substance is not particularly limited, but examples include enzymes such as γ-GTP, pathogenic E. coli such as enteropathogenic E. coli, pathogenic bacteria such as Salmonella and Listeria, actinomycetes, yeast, mold, viruses, and other microorganisms.

Next, the control function 141 places the reagent and magnetic particles into the reaction vessel 50 (step S102). For example, the control function 141 causes the dispensing control device 20 to control the operation of the probe 40, thereby injecting the reagent and magnetic particles from the probe 40 into the reaction vessel 50. Note that step S102 may also be performed manually by the user.

The reagent here includes a detection substance that specifically binds to a specific target substance and a fluorescent dye. For example, an antibody can be used as the detection substance. In this case, the detection substance specifically binds to the target substance, which is an antigen, through an antigen-antibody reaction. Furthermore, any fluorescent substance that emits fluorescence at wavelengths similar to visible light, infrared light, or ultraviolet light can be used as the fluorescent dye. Specific examples of fluorescent dyes include perylene, thioflavin, berberine, fluorescein, rhodamine 123, rhodamine 6G, tetramethylrhodamine, rhodamine B, DiIC12(3), Nile Red, DiIC1(5), and Cy7.

More specifically, the reagent shown in step S102 includes multiple combinations of fluorescent dyes and detection substances that specifically bind to specific target substances. An example of the reagent will be described below using FIG. 3. FIG. 3 is a diagram showing an example of a reagent according to the first embodiment. Note that the following describes the simultaneous detection of ten target substances G1 to G10.

In Figure 3, antibody B1 is a detection substance that specifically binds to target substance G1. Antibody B2 is a detection substance that specifically binds to target substance G2. Antibody B3 is a detection substance that specifically binds to target substance G3. Antibody B4 is a detection substance that specifically binds to target substance G4. Antibody B5 is a detection substance that specifically binds to target substance G5. Antibody B6 is a detection substance that specifically binds to target substance G6. Antibody B7 is a detection substance that specifically binds to target substance G7. Antibody B8 is a detection substance that specifically binds to target substance G8. Antibody B9 is a detection substance that specifically binds to target substance G9. Antibody B10 is a detection substance that specifically binds to target substance G10. Furthermore, fluorescent dyes F1 to F4 are fluorescent dyes that emit fluorescence of different wavelengths.

For example, antibody B1 is included in the reagent in combination with fluorescent dye F1. While the method for combining antibody B1 and fluorescent dye F1 is not particularly limited, one example is a method involving latex particles. Specifically, latex particles are first dispersed in an aqueous medium and mixed with acetone or the like to swell the latex particles. Then, fluorescent dye F1 is added, for example, in the form of an acetone solution. This produces microparticles in which fluorescent dye F1 is encapsulated within the latex particles. Antibody B1 is then added to the liquid containing the microparticles, thereby supporting antibody B1 on the surface of the microparticles. Antibody B1 may be physically adsorbed or chemically bonded to the surface of the microparticles. In this way, fluorescent dye F1 encapsulated within the microparticles and antibody B1 supported on the surface of the microparticles can be combined together. Similarly, antibody B2 is included in the reagent in combination with fluorescent dye F2. Similarly, antibody B3 is included in the reagent in combination with fluorescent dye F3. Similarly, antibody B4 is included in the reagent in combination with fluorescent dye F4.

Furthermore, antibody B5 is included in the reagent in combination with fluorescent dye F1, and also in combination with fluorescent dye F3. For example, the reagent contains microparticles combining antibody B5, fluorescent dye F1, and fluorescent dye F3 via latex particles. Alternatively, the reagent contains both microparticles combining antibody B5 and fluorescent dye F1 via latex particles and microparticles combining antibody B5 and fluorescent dye F3 via latex particles. Similarly, antibody B6 is included in the reagent in combination with fluorescent dye F2, and also in combination with fluorescent dye F4. Similarly, antibody B7 is included in the reagent in combination with fluorescent dye F1, and also in combination with fluorescent dye F2. Similarly, antibody B8 is included in the reagent in combination with fluorescent dye F1, and also in combination with fluorescent dye F4. Similarly, antibody B9 is included in the reagent in combination with fluorescent dye F2, and also in combination with fluorescent dye F3. Similarly, antibody B10 is included in the reagent in combination with fluorescent dye F3, and is also included in the reagent in combination with fluorescent dye F4.

For example, if antibody B1 is the first detection substance, fluorescent dye F1 is an example of a first fluorescent dye, antibody B5 is an example of a second detection substance, and fluorescent dye F3 is an example of a second fluorescent dye. The reagent shown in step S102 is prepared to include at least a combination of the first detection substance and the first fluorescent dye, a combination of the second detection substance and the first fluorescent dye, and a combination of the second detection substance and the second fluorescent dye.

Furthermore, the magnetic particles shown in step S102 include, for example, as shown in FIG. 4, combinations of antibody B1 and magnetic particles, combinations of antibody B2 and magnetic particles, combinations of antibody B3 and magnetic particles, combinations of antibody B4 and magnetic particles, combinations of antibody B5 and magnetic particles, combinations of antibody B6 and magnetic particles, combinations of antibody B7 and magnetic particles, combinations of antibody B8 and magnetic particles, combinations of antibody B9 and magnetic particles, and combinations of antibody B10 and magnetic particles. FIG. 4 is a diagram showing an example of magnetic particles according to the first embodiment.

For example, in step S102, the reagent shown in FIG. 3 and the magnetic particles shown in FIG. 4 are injected into the reaction vessel 50 at approximately the same time and mixed with the test sample P. Here, specific binding occurs depending on the substance contained in the test sample P. For example, if the test sample P contains a target substance G1, microparticles combining antibody B1 and fluorescent dye F1 will bind to the target substance G1, and magnetic particles combined with antibody B1 will also bind.

Next, the control function 141 controls the magnetic field generator 32 to attract the magnetic particles with magnetic force (step S103). For example, if the magnetic field generator 32 is an electromagnet, the control function 141 can generate magnetic force by passing a current through the electromagnet, thereby attracting the magnetic particles to the side of the reaction vessel 50. Alternatively, if the magnetic field generator 32 is a permanent magnet, the control function 141 can attract the magnetic particles to the side of the reaction vessel 50 by bringing the permanent magnet closer to the reaction vessel 50.

Next, the control function 141 drains the liquid in the reaction vessel 50 (step S104). Here, the magnetic particles attracted by the magnetic force shown in step S103 remain in the reaction vessel 50. In addition, the target substance bound to the magnetic particles, and the detection substance and fluorescent dye bound to the target substance also remain in the reaction vessel 50. In addition, the control function 141 adds a buffer to the reaction vessel 50 (step S105).
The control function 141 can remove the detection substance and fluorescent dye that have not bound to the target substance through the processing of steps S102 to S105.

Next, the control function 141 irradiates light from the light source 31 onto the sample in the reaction vessel 50 (step S106). That is, the control function 141 irradiates light onto a mixed sample obtained by mixing a test sample with a reagent containing multiple combinations of fluorescent dyes and detection substances that specifically bind to specific target substances. Here, the fluorescent dyes contained in the mixed sample are excited, and fluorescence of a wavelength corresponding to the type of fluorescent dye is emitted.

After stopping the irradiation of light from the light source 31, the detection function 142 controls the operation of the photodetector 33 to detect the fluorescence emitted from the mixed sample in the reaction vessel 50 for each wavelength band (step S107). For example, if the photodetector 33 is an MSI camera, the detection function 142 acquires an image for each wavelength band as the detection result of the fluorescence for each wavelength band. In other words, the detection function 142 detects the fluorescence for each wavelength band using multispectral imaging, which acquires an image for each wavelength band.

An example of fluorescence detection processing by the detection function 142 will now be described using FIG. 5. For example, as shown in FIG. 5, the detection function 142 sets four wavelength bands, windows W1 to W4. Here, window W1 corresponds to the wavelength of fluorescence emitted from fluorescent dye F1. For example, window W1 includes the peak wavelength of fluorescence emitted from fluorescent dye F1. Window W2 corresponds to the wavelength of fluorescence emitted from fluorescent dye F2. Window W3 corresponds to the wavelength of fluorescence emitted from fluorescent dye F3. Window W4 corresponds to the wavelength of fluorescence emitted from fluorescent dye F4. Windows W1 to W4 may be wavelength bands corresponding to visible light, or may be wavelength bands corresponding to ultraviolet or infrared light. Note that FIG. 5 is a diagram for explaining fluorescence detection processing for each wavelength band according to the first embodiment.

The fluorescence detection results by the detection function 142 can be shown as the correspondence between the wavelength of the fluorescence and the output of the photodetector 33 (detector output), as shown in the lower diagram of Figure 6, for example. Figure 6 is a diagram showing an example of a fluorescence detection result according to the first embodiment. For example, if the photodetector 33 is an MSI camera, the output of the photodetector 33 is the signal intensity in each image acquired for each wavelength band. For example, the detection function 142 compares the output of the photodetector 33 with a threshold value for each of windows W1 to W4 and performs binarization. For example, in Figure 6, when the output of the photodetector 33 exceeds the threshold value, it is marked with an "O", and when the output of the photodetector 33 is equal to or less than the threshold value, it is marked with an "X".

In Figure 6, windows W1 and W3 are marked with "○". That is, Figure 6 shows a case where fluorescence in the wavelength bands of windows W1 and W3 is detected. Next, the identification function 143 identifies the target substance contained in the test sample P based on the combination of multiple wavelength bands in which fluorescence is detected (step S108).

For example, if fluorescence in the wavelength bands of window W1 and window W3 is detected, it can be inferred that the mixed sample contains fluorescent dye F1 corresponding to window W1 and fluorescent dye F3 corresponding to window W3. Furthermore, as shown in FIG. 3, antibody B5 is combined with both fluorescent dye F1 and fluorescent dye F3. From the above, identification function 143 can identify antibody B5 as the detection substance that binds to the target substance in the mixed sample. In other words, identification function 143 can identify target substance G5 that specifically binds to antibody B5 as the target substance contained in test sample P in the case shown in FIG. 6.

As described above, the identification function 143 can identify the target substance contained in the test sample P by identifying the detection substance bound to the target substance in the mixed sample based on the combination of multiple wavelength bands in which fluorescence was detected. For example, as shown in FIG. 7, the identification function 143 may generate in advance a correspondence table that associates wavelength band combinations such as windows W1 to W4 with standard substances and store the table in memory 11. In this case, the identification function 143 can identify the target substance contained in the test sample P by comparing the fluorescence detection results obtained by the detection function 142 with the correspondence table. That is, the memory 11 stores data on the correspondence between the target substance and the detected wavelength information. The correspondence data associates multiple wavelength information with at least one target substance. The identification function 143 can then identify the target substance contained in the test sample P using the correspondence data. FIG. 7 is a diagram showing an example of a correspondence table between wavelength band combinations and standard substances according to the first embodiment.

Alternatively, the identification function 143 can identify a target substance using the table shown in FIG. 3 as correspondence data. That is, the memory 11 stores correspondence data between target substances and types of detection substances. The correspondence data associates multiple detection substances with at least one target substance. The identification function 143 then uses the correspondence data to identify the target substance contained in the test sample P. For example, if fluorescence in the wavelength bands of windows W1 and W3 is detected as shown in FIG. 6, the identification function 143 can estimate that the mixed sample contains fluorescent dye F1 corresponding to window W1 and fluorescent dye F3 corresponding to window W3. Furthermore, the identification function 143 can estimate that the mixed sample contains (a) antibodies B1, B5, B7, and B8 bound to fluorescent dye F1, and (b) antibodies B3, B5, B9, and B10 bound to fluorescent dye F3. The identification function 143 can then use the table in Figure 3 to identify target substance G5 that satisfies (a) and (b).

Then, the output function 144 outputs the test results (step S109). For example, the output function 144 displays the target substance identified by the identification function 143 on the display 13 as the test results. Furthermore, if fluorescence is detected using multispectral imaging, the output function 144 may display images acquired for each wavelength band along with the identified target substance.

Note that while FIG. 1 illustrates the inspection device 1 as being equipped with a display 13, the inspection device 1 may also be equipped with a projector instead of the display 13. The projector can project onto a screen, wall, floor, etc. under the control of the output function 144. That is, the output function 144 may notify the user of the inspection results by projection from the projector. The output function 144 may also print out the inspection results. The output function 144 may also notify the user of the inspection results by audio, etc. The output function 144 may also send the inspection results to an external server for storage. For example, the output function 144 registers the evaluation results on a system such as an HIS (Hospital Information System). In this case, the user can access the system at will and refer to the evaluation results.

As described above, according to the first embodiment, the control function 141 irradiates light onto a mixed sample obtained by mixing a test sample with a reagent containing multiple combinations of fluorescent dyes and detection substances that specifically bind to specific target substances. The detection function 142 detects the fluorescence emitted from the mixed sample in response to the light irradiation for each wavelength band. The identification function 143 identifies the target substance contained in the test sample P based on the combination of multiple wavelength bands in which the fluorescence was detected. This allows the testing device 1 to simultaneously detect ten types of target substances, G1 to G10, as shown in FIG. 7, for example.

Another possible method for simultaneously detecting target substances G1 to G10 is to use 10 types of fluorescent dyes. That is, a test could be performed using a reagent in which different types of fluorescent dyes are combined with antibodies B1 to B10, and the target substances G1 to G10 could be simultaneously detected depending on whether or not fluorescence corresponding to each fluorescent dye is detected. However, in this case, it would be necessary to set up 10 windows corresponding to the 10 types of fluorescent dyes to detect the fluorescence.

When multiple windows are used to detect fluorescence, the width of each individual window becomes narrow, which can reduce sensitivity and S/N in detecting fluorescence. Furthermore, performing detection for each of the multiple windows can complicate the system, potentially increasing the cost of testing.

In contrast, the inspection device 1 according to the first embodiment can detect fluorescence using, for example, four windows, allowing for simultaneous detection of 10 types of target substances. In other words, the inspection device 1 is capable of simultaneously detecting multiple types of target substances while reducing the reduction in sensitivity and S/N caused by narrowing the window width, thereby improving the detection accuracy of target substances.

The number of target substances that can be simultaneously detected by the inspection device 1 varies depending on the number of fluorescent dyes used. For example, when N types of fluorescent dyes (N is a natural number of 2 or more) are used, the inspection device 1 can simultaneously detect "N + _N C ₂ " target substances.

Second Embodiment
In the second embodiment, a case where the detection of a target substance described in the first embodiment is performed quantitatively will be described. The testing device 1 according to the second embodiment has a configuration similar to that of the testing device 1 shown in FIG. 1, but differs in that it has a reaction vessel 51 shown in FIG. 8A instead of the reaction vessel 50. FIG. 8A is a diagram showing an example of a reaction vessel according to the second embodiment. Hereinafter, the same reference numerals will be used to designate the points described in the first embodiment, and description thereof will be omitted.

The reaction vessel 51 shown in Figure 8A is a plate provided with multiple cells (places for holding measurement liquid), such as cell C1, cell C2, cell C3, and cell C4. The control function 141 can individually dispense the test sample P and reagents into each of cell C1, cell C2, cell C3, and cell C4.

Here, we will explain the reagent to be dispensed into each of the multiple cells using Figures 8B and 8C. Figures 8B and 8C are diagrams showing an example of a reagent according to the second embodiment.

As shown in Figure 8B, the reagents include a combination of antibody B1 and fluorescent dye F1, a combination of antibody B2 and fluorescent dye F2, a combination of antibody B3 and fluorescent dye F3, a combination of antibody B4 and fluorescent dye F4, a combination of antibody B5 and fluorescent dye F1, a combination of antibody B5 and fluorescent dye F3, a combination of antibody B6 and fluorescent dye F2, and a combination of antibody B6 and fluorescent dye F4. Note that antibody B1 is a detection substance that specifically binds to target substance G1. Antibody B2 is a detection substance that specifically binds to target substance G2. Antibody B3 is a detection substance that specifically binds to target substance G3. Antibody B4 is a detection substance that specifically binds to target substance G4. Antibody B5 is a detection substance that specifically binds to target substance G5. Antibody B6 is a detection substance that specifically binds to target substance G6. Hereinafter, as shown in Figure 8C, the combination of antibody B1 and fluorescent dye F1 will also be referred to as reagent R1. The combination of antibody B2 and fluorescent dye F2 is also referred to as reagent R2. The combination of antibody B3 and fluorescent dye F3 is also referred to as reagent R3. The combination of antibody B4 and fluorescent dye F4 is also referred to as reagent R4. The combination of antibody B5 and fluorescent dye F1 is also referred to as reagent R5. The combination of antibody B5 and fluorescent dye F3 is also referred to as reagent R6. The combination of antibody B6 and fluorescent dye F2 is also referred to as reagent R7. The combination of antibody B6 and fluorescent dye F4 is also referred to as reagent R8.

The control function 141 dispenses test sample P into each of cells C1 to C4, as well as a reagent containing reagents R1 to R8. For example, the control function 141 dispenses the same amount of test sample P into each cell. The control function 141 also dispenses reagents R1 to R8 and diluent while changing the dispensed amount so that the reagent concentration varies for each cell. For example, as shown in Figure 8D, the control function 141 dispenses reagents R1 to R8 and diluent into each cell while changing the concentration for each cell. For example, the control function 141 dispenses each reagent and diluent into cell C1 so that each of reagents R1 to R8 is 0.5%. Note that Figure 8D is a diagram showing an example of a reagent according to the second embodiment.

For example, the control function 141 first prepares a reagent containing 0.5% of each of the reagents R1 to R8. For example, each of the reagents R1 to R8 is a microparticle formed by bonding a fluorescent dye and an antibody via latex particles. The control function 141 prepares the reagent so that the mass percentage concentration of each microparticle is 0.5%. That is, the control function 141 mixes the reagents R1 to R8 with a diluent to prepare a reagent containing 0.5% microparticles corresponding to the reagent R1, 0.5% microparticles corresponding to the reagent R2, 0.5% microparticles corresponding to the reagent R3, 0.5% microparticles corresponding to the reagent R4, 0.5% microparticles corresponding to the reagent R5, 0.5% microparticles corresponding to the reagent R6, 0.5% microparticles corresponding to the reagent R7, and 0.5% microparticles corresponding to the reagent R8. The control function 141 then dispenses the prepared reagent into the cell C1. Similarly, the control function 141 dispenses 1.0% of each of the reagents R1 to R8 into cell C2. Similarly, the control function 141 dispenses 1.5% of each of the reagents R1 to R8 into cell C3. Similarly, the control function 141 dispenses 2.0% of each of the reagents R1 to R8 into cell C4.

Next, detection function 142 detects the fluorescence emitted from each of cells C1 to C4 for each wavelength band. That is, detection function 142 detects the fluorescence emitted from multiple mixed samples with varying reagent concentrations for each wavelength band and each reagent concentration. Here, detection function 142 may detect the fluorescence from each cell in parallel or sequentially. For example, detection function 142 sets four wavelength bands, windows W1 to W4, as in Figure 5. Furthermore, detection function 142 compares the output of photodetector 33 with a threshold for each of windows W1 to W4 and binarizes it into "○" or "×."

Figure 8E shows an example of fluorescence detection results according to the second embodiment. In Figure 8E, all windows in cells C1 and C2 are marked with an "X," indicating that the output of the photodetector 33 did not exceed the threshold in any wavelength band. Also, in Figure 8E, window W3 in cells C3 and C4 is marked with an "O," indicating that the output of the photodetector 33 exceeded the threshold in window W3.

Here, the identification function 143 can identify the target substance contained in the test sample P based on the combination of multiple wavelength bands in which fluorescence was detected. For example, the identification function 143 can identify the target substance using the table of FIG. 8B as correspondence data. That is, in the case shown in FIG. 8D, because fluorescence in the wavelength band of window W3 is detected, it can be inferred that the fluorescent dye F3 corresponding to window W3 is contained in the test sample P. Furthermore, because fluorescence in the wavelength band of window W1 is not detected, the identification function 143 can infer that the fluorescent dye F1 corresponding to window W1 is not contained in the test sample P. From the above, the identification function 143 can identify the target substance G3 contained in the test sample P.

Furthermore, the identification function 143 can calculate the amount of target substance contained in the test sample P. Specifically, in Figure 8E, the window W3 is marked "X" in cells C1 and C2 even though the test sample P contains target substance G3. This is because the concentration of reagent R3 in cells C1 and C2 is low, resulting in a low amount of luminescence and preventing the output of the photodetector 33 from exceeding the threshold. In other words, in cells C1 and C2, the concentration of reagent R3 is low relative to the amount of target substance G3 contained in the test sample P, and the amount of luminescence is low and dependent on the amount of reagent R3. As the amount of reagent R3 increases, the amount of luminescence also increases, and in cells C3 and C4, the output of the photodetector 33 exceeds the threshold.

Here, if the amount of reagent R3 is gradually increased, eventually the amount of reagent R3 will be sufficient for the amount of target substance G3, and further increases in the amount of reagent R3 will no longer increase the amount of luminescence. In other words, the identification function 143 can determine the value at which the amount of luminescence saturates for the amount of reagent R3. Furthermore, the value at which the amount of luminescence saturates depends on the amount of target substance G3 contained in the test sample P.

For example, if the photodetector 33 is an MSI camera, the detection function 142 causes the MSI camera to capture an image including cells C1 to C4. Here, the MSI camera divides the wavelength of light into multiple wavelength bands and acquires an image for each wavelength band. For example, the MSI camera acquires two-dimensional images for each of the four wavelength bands of windows W1 to W4. The detection function 142 also analyzes the two-dimensional images acquired by the MSI camera to determine the amount of light emitted by each cell. For example, the detection function 142 sets regions of interest corresponding to cells C1 to C4 in the two-dimensional images for each wavelength band acquired by the MSI camera, and determines the amount of light emitted by each cell based on the signal intensity within the regions of interest. For example, the detection function 142 can determine the amount of light emitted in the wavelength band of window W3 in cell C3 based on the signal intensity within a region of interest corresponding to cell C3, which is set in the two-dimensional image of window W3 acquired by the MSI camera.

For example, in Figure 8E, if the amount of light emitted in the wavelength band of window W3 does not change between cell C3 and cell C4, the identification function 143 can determine that the amount of light emitted has saturated at 1.5%. The identification function 143 can then calculate the amount or concentration of target substance G3 contained in the test sample P based on the value at which the amount of light emitted saturates. In other words, the identification function 143 can calculate the amount or concentration of target substance G3 contained in the test sample P based on the amount of light emitted for each reagent concentration in wavelength band W3 in which fluorescence was detected.

(Third embodiment)
Although the first and second embodiments have been described above, the present invention may be embodied in various different forms other than the above-described embodiments.

For example, in the above-described embodiment, the target substance and the detection substance are described as specifically binding to each other. However, depending on the selection of the target substance and the detection substance, non-specific binding may occur along with specific binding. Therefore, the identification function 143 may identify the standard substance while taking non-specific binding into consideration.

The process of identifying a reference substance that takes non-specific binding into consideration will be explained below with reference to Figure 9. Figure 9 shows an example of a fluorescence detection result according to the third embodiment. In Figure 9, target substances G3 and G4 have similar structures, and the explanation will be given assuming that not only does antibody B4 specifically bind to target substance G4, but antibody B3 also binds non-specifically.

Figure 9 shows the results of fluorescence detection using the reagent shown in Figure 3 when the test sample P contains target substance G4. That is, in Figure 9, antibody B4 specifically binds to target substance G4, causing fluorescence emitted from fluorescent dye F4 to be detected, and the output of photodetector 33 is high in window W4. Also in Figure 9, antibody B3 nonspecifically binds to target substance G4, causing fluorescence emitted from fluorescent dye F3 to be detected, and the output of photodetector 33 is also high in window W3, although not as high as in window W4.

In such a case, the identification function 143 identifies the target substance contained in the test sample P based on the spectrum indicating the fluorescence detection results. For example, the identification function 143 identifies the detection substance that specifically binds to the target substance in the mixed sample and the detection substance that non-specifically binds to the target substance based on the detector output waveform shown in the lower diagram of Figure 9, thereby identifying the target substance contained in the test sample P. In other words, although fluorescence is detected in both window W3 and window W4, the identification function 143 determines that antibody B3 binds non-specifically and antibody B4 binds specifically, and that the target substance contained in the test sample P is target substance G4.

For example, a correspondence table that associates the detector output waveform with the target substance is generated in advance and stored in memory 11. In this case, the identification function 143 can identify the target substance contained in the test sample P by comparing the spectrum indicating the fluorescence detection results with the correspondence table.

As another example, the identification function 143 can identify target substances contained in the test sample P using machine learning techniques. For example, by performing a test using a known target substance, it is possible to obtain a large number of combinations of known target substances and spectra showing fluorescence detection results. Furthermore, a machine learning algorithm such as a neural network can be trained using the spectrum showing fluorescence detection results as input data and the known target substance as output data to obtain a trained model. Such a trained model is configured to accept input of a spectrum showing fluorescence detection results and identify the target substance contained in the test sample P.

Although the photodetector 33 has been described as an MSI camera, the embodiment is not limited to this. For example, the photodetector 33 may be a device that detects fluorescence in each wavelength band by spectrally separating the fluorescence. For example, the photodetector 33 includes a spectral device such as a grating (diffraction grating) or a prism, and a light-receiving device such as a CCD (charge-coupled device) that receives the dispersed fluorescence. In this case, the fluorescence incident on the spectral device is spectrally separated according to wavelength and irradiated onto one of the light-receiving devices installed in multiple positions. The detection function 142 can then detect the fluorescence in each wavelength band depending on which light-receiving device receives the light. Additionally, the detection function 142 can detect the fluorescence in each wavelength band using various techniques. For example, the detection function 142 can detect the fluorescence in each wavelength band using known techniques that use secondary antibodies or microarrays.

Although Figure 3 illustrates the detection of 10 types of target substances and Figure 8B illustrates the detection of 6 types of target substances, the number of target substances to be detected can be changed as desired. Furthermore, detection is not limited to cases where multiple types of target substances are detected; the target substance to be detected may be one type. For example, the identification function 143 may determine the presence or absence of one target substance on the condition that fluorescence is detected in multiple wavelength bands.

The term "processor" used in the above description refers to circuits such as a CPU, a GPU (Graphics Processing Unit), an Application Specific Integrated Circuit (ASIC), a programmable logic device (e.g., a Simple Programmable Logic Device (SPLD), a Complex Programmable Logic Device (CPLD), and a Field Programmable Gate Array (FPGA)). When the processor is, for example, a CPU, the processor realizes its functions by reading and executing programs stored in a memory circuit. On the other hand, if the processor is, for example, an ASIC, instead of storing the program in a memory circuit, the function is directly incorporated into the processor circuit as a logic circuit. Note that each processor in the embodiments is not limited to being configured as a single circuit, but may be configured as a single processor by combining multiple independent circuits to realize its function. Furthermore, multiple components in each diagram may be integrated into a single processor to realize its function.

Furthermore, in FIG. 1, a single memory 11 is described as storing programs corresponding to each processing function of the processing circuit 14. However, the embodiment is not limited to this. For example, multiple memories 11 may be distributed and the processing circuit 14 may read corresponding programs from individual memories 11. Furthermore, instead of storing programs in memory 11, the programs may be directly embedded in the processor circuitry. In this case, the processor realizes the functions by reading and executing the programs embedded in the circuitry.

The components of each device according to the above-described embodiments are conceptual and functional, and do not necessarily need to be physically configured as shown in the illustrations. In other words, the specific form of distribution and integration of each device is not limited to that shown, and all or part of the devices can be functionally or physically distributed and integrated in any unit depending on various loads and usage conditions. Furthermore, all or any part of the processing functions performed by each device can be realized by a CPU and a program analyzed and executed by the CPU, or can be realized as hardware using wired logic.

The inspection method described in the above-mentioned embodiments can be realized by executing a pre-prepared program on a computer such as a personal computer or workstation. This program can be distributed via a network such as the Internet. This program can also be recorded on a non-transitory computer-readable recording medium such as a hard disk, flexible disk (FD), CD-ROM, MO, or DVD, and executed by being read from the recording medium by a computer.

At least one of the embodiments described above makes it possible to simultaneously detect multiple types of target substances with high accuracy.

While several embodiments have been described, these embodiments are presented as examples and are not intended to limit the scope of the invention. These embodiments may be embodied in a variety of other forms, and various omissions, substitutions, modifications, and combinations of embodiments may be made without departing from the spirit of the invention. These embodiments and their variations are included within the scope and spirit of the invention, as well as within the scope of the invention and its equivalents as set forth in the claims.

1 Inspection device 10 Console device 14 Processing circuit 141 Control function 142 Detection function 143 Identification function 144 Output function

Claims (8)

a detection unit for detecting, for each wavelength band, fluorescence emitted from a mixed sample obtained by mixing a sample with a reagent containing multiple combinations of a detection substance that specifically binds to a specific target substance and a fluorescent dye;
an identification unit that identifies the target substance contained in the sample based on a combination of a plurality of wavelength bands in which the fluorescence is detected,
the detection unit acquires, by multispectral imaging , a two-dimensional image having two axes in spatial directions for each of a plurality of wavelength bands , and detects the fluorescence for each wavelength band for each of a plurality of regions set in the two-dimensional image for each of the wavelength bands;
The identification unit identifies the target substance for each of the plurality of regions. The inspection device described in claim 1, wherein the identification unit determines the presence or absence of a single target substance based on the condition that fluorescence is detected in multiple wavelength bands. The testing device according to claim 1 or 2, wherein the reagents include at least a combination of a first detection substance and a first fluorescent dye, a combination of a second detection substance and a first fluorescent dye, and a combination of a second detection substance and a second fluorescent dye. The testing device according to any one of claims 1 to 3, wherein the identification unit identifies the detection substance bound to the target substance in the mixed sample based on a combination of multiple wavelength bands in which the fluorescence is detected, thereby identifying the target substance contained in the sample. The inspection device described in any one of claims 1 to 4, wherein the identification unit identifies the target substance contained in the sample based on a spectrum indicating the fluorescence detection results. a storage unit that stores data on a correspondence relationship between the target substance and the type of the detection substance or wavelength information to be detected, in which a plurality of detection substances or wavelength information are associated with at least one target substance as the correspondence relationship data;
6. The inspection device according to claim 1, wherein the identifying unit identifies the target substance using the correspondence data. the detection unit detects fluorescence emitted from a plurality of mixed samples in which the concentrations of the reagent are varied, for each wavelength band and for each concentration of the reagent;
7. The testing device according to claim 1, wherein the identification unit further calculates the amount or concentration of the target substance contained in the sample based on the amount of light emitted for each concentration of the reagent in the wavelength band in which the fluorescence is detected. A reagent containing multiple combinations of a detection substance and a fluorescent dye that specifically binds to a specific target substance is mixed with the sample, and the fluorescence emitted from the mixed sample is detected for each wavelength band;
identifying the target substance contained in the sample based on a combination of a plurality of wavelength bands in which the fluorescence is detected;
By multispectral imaging , a two-dimensional image having two axes in the spatial direction is acquired for each of a plurality of wavelength bands , and the fluorescence is detected for each wavelength band in each of a plurality of regions set in the two-dimensional image for each of the wavelength bands;
An inspection method, comprising identifying the target substance for each of the plurality of regions.